Aminoadipate-semialdehyde synthase inhibitors and methods of treating cancer

Abstract

This disclosure relates to methods of treating cancer comprising administering an effective amount of an aminoadipate-semialdehyde synthase inhibitor to a subject in need thereof. In certain embodiments, the aminoadipate-semialdehyde synthase inhibitor is N-benzyl-N,N-dimethylhexadecan-1-aminium chloride (cetalkonium chloride) or salts thereof. In certain embodiments, this disclosure relates to pharmaceutical compositions comprising an aminoadipate-semialdehyde synthase inhibitor such as N-benzyl-N,N-dimethylhexadecan-1-aminium chloride (cetalkonium chloride) or salts thereof.

Description

[0001] AMINOADIPATE-SEMIALDEHYDE SYNTHASE INHIBITORS AND METHODS OF TREATING CANCER

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Application No. 63 / 461,451 filed April 24, 2023. The entirety of this application is hereby incorporated by reference for all purposes.

[0004] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0005] This invention was made with government support under CA266613 awarded by the National Institutes of Health. The government has certain rights in the invention.

[0006] BACKGROUND

[0007] Advanced cancer is typically used to describe cancers that quick progress and are not usually curable with treatment but are sometimes controlled over a period. Advanced cancers can be characterized as locally advanced or metastatic. Locally advanced refers to some extensions from the original location but has not yet spread to other body parts. “Metastatic cancer” refers to a cancer that has traveled, e.g., through the bloodstream or the lymph system, from a primary site to other tissues of the body where the migrated cancer cells establish in a remote area(s). Metastatic cancer is named after the tissue from which it originated. For example, pancreatic cancer that has spread to the lungs is called “metastatic pancreatic cancer to the lungs.” The use of modern chemotherapy agents to treat cancer have improved survival duration. However, treatment effectiveness is not always universal; and remission is common in various situations. Thus, improved methods of managing cancer therapies are needed.

[0008] Mamoor reports differential expression of aminoadipate-semialdehyde synthase in cancers of the breast. OSF Preprints, 2021.

[0009] Wissing et al. report small-molecule screening of PC3 prostate cancer cells identifies tilorone dihydrochloride to selectively inhibit cell growth based on cyclin-dependent kinase 5 expression. Oncol Rep. 2014, 32(1): 419-424.

[0010] Duell et al. report benefits of cetalkonium chloride cationic oil-in-water nanoemulsions for topical ophthalmic drug delivery. J Pharm Pharmacol, 2014, 66(4): 531-541. References cited herein are not an admission of prior art.

[0011] SUMMARY

[0012] This disclosure relates to methods of treating cancer comprising administering an effective amount of an aminoadipate-semialdehyde synthase inhibitor to a subject in need thereof. In certain embodiments, the aminoadipate-semialdehyde synthase inhibitor is N-benzyl-N,N- dimethylhexadecan-l-aminium chloride (cetalkonium chloride) or salts thereof. In certain embodiments, this disclosure relates to pharmaceutical compositions comprising an aminoadipate- semialdehyde synthase inhibitor such as N-benzyl-N,N-dimethylhexadecan-l-aminium chloride (cetalkonium chloride) or salts thereof.

[0013] In certain embodiments, methods disclosed herein further comprise administering an additional anti-cancer agent in combination with the aminoadipate-semialdehyde synthase inhibitor such as N-benzyl-N,N-dimethylhexadecan-l-aminium chloride (cetalkonium chloride) or alternative salts thereof.

[0014] In certain embodiments, the aminoadipate-semialdehyde synthase inhibitor is a N-benzyl- N,N-dimethylhexadecan-l-aminium salt, prodrug, metabolite, or derivative thereof. In certain embodiments, the aminoadipate-semialdehyde synthase inhibitor is N-benzyl-N,N- dimethylhexadecan-l-aminium chloride (cetalkonium chloride).

[0015] In certain embodiments, the aminoadipate-semialdehyde synthase inhibitor is an antibody, small molecule compound, peptide, or siRNA.

[0016] In certain embodiments, the cancer is selected from a solid cancer, advanced cancer, metastatic cancer, and / or hematological cancer. In certain embodiments, the solid cancer has metastasized or is a locally advanced cancer. In certain embodiments, the solid cancer is selected from liver cancer, lung cancer, ovarian cancer, bladder cancer, lung cancer, breast cancer, colon cancer, rectal cancer, endometrial cancer, pancreatic cancer, kidney cancer, prostate cancer, thyroid cancer, or brain cancer. In certain embodiments, hematological cancer is leukemia, lymphoma, or multiple myeloma.

[0017] In certain embodiments, methods further comprise administering an additional anti-cancer agent.

[0018] In certain embodiments, this disclosure relates to pharmaceutical composition comprising an aminoadipate-semialdehyde synthase inhibitor and a pharmaceutically acceptable excipient. In certain embodiments, the aminoadipate-semialdehyde synthase inhibitor is a N-benzyl-N,N- dimethylhexadecan-l-aminium salt, prodrug, metabolite, or derivative thereof. In certain embodiments, the aminoadipate-semialdehyde synthase inhibitor is N-benzyl-N,N- dimethylhexadecan-l-aminium chloride (cetalkonium chloride).

[0019] In certain embodiments, the pharmaceutical composition is in the form of a lotion, gel, cream, pill, tablet, capsule, or gel capsule. In certain embodiments, the pharmaceutical composition is in the form of a sterilized pH buffered aqueous salt solution or a saline phosphate buffer between a pH of 6 to 8, optionally comprising a saccharide or polysaccharide.

[0020] In certain embodiments, this disclosure relates to methods of treating cancer comprising administering an effective amount of an aminoadipate-semialdehyde synthase inhibitor to a subject in need thereof. In certain embodiments, this disclosure relates to methods of detecting amounts of aminoadipate-semialdehyde synthase in a sample to determine or monitor whether the subject is sensitive or resistant to an aminoadipate-semialdehyde synthase inhibitor or combination of chemotherapy agents comprising the same.

[0021] In certain embodiments, this disclosure relates to methods of detecting amounts of aminoadipate-semialdehyde synthase in a sample to determine whether the subject is sensitive or resistant or is likely to develop resistance to a chemotherapy agent or combination of chemotherapy agents comprising the same. In certain embodiments, the sample is derived from tumor cells or cell-free DNA contained in a blood sample of a subject diagnosed with cancer.

[0022] In certain embodiments, this disclosure relates to medicaments comprising an aminoadipate-semialdehyde synthase inhibitor for use in treating cancer as provided herein. In certain embodiments, this disclosure relates to medicaments comprising an aminoadipate- semialdehyde synthase inhibitor for use in treating cancer as provided herein.

[0023] BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

[0024] Figure 1A shows data on the ability of N-benzyl-N,N-dimethylhexadecan-l-aminium chloride (Cetalkonium chloride, CKC) to inhibit Aminoadipate-Semialdehyde Synthase (AASS).

[0025] Figure IB shows data on the ability of CKC to reduce invasiveness of Hl 299 cells (human non-small cell lung cancer cell).

[0026] Figure 1C shows data on the ability of CKC to reduce invasiveness of 37B cells (human metastatic squamous cell carcinoma larynx cell). Figure ID shows data on the ability of CKC to reduce invasiveness of MDA-MB-231 cells (human breast cancer cell).

[0027] Figure IE shows data on the ability of CKC to reduce invasiveness of A2780 cells (human ovarian cancer cell line).

[0028] Figure 2A shows data on the ability of CKC to dose dependently reduce metastasis tumor volume when administered to mice in vivo.

[0029] Figure 2B shows data on the ability of CKC to dose dependently reduce the number of mice showing liver metastasis when administered to mice in vivo.

[0030] Figure 2C shows data on the ability of CKC to dose dependently reduce the number of mice showing lung metastasis when administered to mice in vivo.

[0031] Figure 2D shows data on the ability of CKC to dose dependently reduce the number of mice showing liver metastasis when administered to mice in vivo.

[0032] Figure 3 shows data indicating CKC attenuates cell invasion through targeting AASS.

[0033] Figure 4 shows data using CKC in a Xenograft mouse model (patient-derived ovarian cancer).

[0034] DETAILED DISCUSSION

[0035] Before the present disclosure is described in greater detail, it is to be understood that this disclosure is not limited to particular embodiments described, and as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims.

[0036] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, the preferred methods and materials are now described.

[0037] All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference and are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present disclosure is not entitled to antedate such publication by virtue of prior disclosure. Further, the dates of publication provided could be different from the actual publication dates that may need to be independently confirmed.

[0038] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present disclosure. Any recited method can be carried out in the order of events recited or in any other order that is logically possible.

[0039] An "embodiment" is an example, and not necessarily limited to such example. Embodiments of the present disclosure will employ, unless otherwise indicated, techniques of medicine, organic chemistry, biochemistry, molecular biology, pharmacology, and the like, which are within the skill of the art. Such techniques are explained fully in the literature.

[0040] It must be noted that, as used in the specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. In this specification and in the claims that follow, reference will be made to a number of terms that shall be defined to have the following meanings unless a contrary intention is apparent.

[0041] Reference to “aminoadipate-semialdehyde synthase” protein refers to the alphaaminoadipic semialdehyde synthase, mitochondrial [Homo sapiens] with GenBank NCBI Reference Sequence: NP_ 005754. The N-terminal and the C-terminal portions of this enzyme contain lysine-ketoglutarate reductase and saccharopine dehydrogenase activity, respectively, resulting in the conversion of lysine to alpha-aminoadipic semialdehyde. An inhibitor refers to a specific binding agent for lysine-ketoglutarate reductase or saccharopine dehydrogenase that has the effect of decreasing alpha-aminoadipic semialdehyde production from lysine in vivo or in vitro.

[0042] A “specific binding agent” may be a protein, peptide, nucleic acid, carbohydrate, lipid, or small molecular weight compound that specifically binds to the aminoadipate-semialdehyde synthase protein. In certain embodiments, the specific binding agent according to the present disclosure is an antibody or binding fragment thereof (e.g., Fab, F(ab')2), peptide or a peptibody. WOOO / 24782 and W003 / 057134 (incorporated herein by reference) describe and teach making binding agents that contain a randomly generated peptide which binds a desired target. A specific binding agent can be a proteinaceous polymeric molecule (a “large molecule”) such as an antibody or Fc-peptide fusion, or a non-proteinaceous non-polymeric molecule typically having a molecular weight of less than about 1200 Daltons (a “small molecule”).

[0043] As used herein, "subject" refers to any animal, preferably a human patient, livestock, or domestic pet.

[0044] As used herein, the terms "treat" and "treating" are not limited to the case where the subject (e.g., patient) is cured and the disease is eradicated. Rather, embodiments, of the present disclosure also contemplate treatment that merely reduces symptoms, and / or delays disease progression.

[0045] As used herein, the term "combination with" when used to describe administration with an additional treatment means that the agent may be administered prior to, together with, or after the additional treatment, or a combination thereof.

[0046] The term "effective amount" refers to that amount of a compound or pharmaceutical composition described herein that is sufficient to effect the intended application including, but not limited to, disease treatment, as illustrated below. The therapeutically effective amount can vary depending upon the intended application (in vitro or in vivo), or the subject and disease condition being treated, e.g., the weight and age of the subject, the severity of the disease condition, the manner of administration and the like, which can readily be determined by one of ordinary skill in the art. The specific dose will vary depending on, for example, the particular agent chosen, the dosing regimen to be followed, whether it is administered in combination with other agents, timing of administration, the tissue to which it is administered, and the physical delivery system in which it is carried.

[0047] "Cancer" refers any of various cellular diseases with malignant neoplasms characterized by the proliferation of cells. It is not intended that the diseased cells must actually invade surrounding tissue and metastasize to new body sites. Cancer can involve any tissue of the body and have many different forms in each body area. Within the context of certain embodiments, whether "cancer is reduced" may be identified by a variety of diagnostic manners known to one skill in the art including, but not limited to, observation of the reduction in size or number of tumor masses or if an increase of apoptosis of cancer cells observed, e.g., if more than a 5 % increase in apoptosis of cancer cells is observed for a sample compound compared to a control without the compound. It may also be identified by a change in relevant biomarker or gene expression profile, such as PSA for prostate cancer, HER2 for breast cancer, or others. The cancer to be treated in the context of the present disclosure may be any type of cancer or tumor. These tumors or cancer include, and are not limited to, tumors of the hematopoietic and lymphoid tissues or hematopoietic and lymphoid malignancies, tumors that affect the blood, bone marrow, lymph, and lymphatic system. Hematological malignancies may be derive from either of the two major blood cell lineages: myeloid and lymphoid cell lines. The myeloid cell line normally produces granulocytes, erythrocytes, thrombocytes, macrophages, and mast cells; the lymphoid cell line produces B, T, NK and plasma cells. Lymphomas, lymphocytic leukemias, and myeloma are from the lymphoid line, while acute and chronic myelogenous leukemia, myelodysplastic syndromes and myeloproliferative diseases are myeloid in origin.

[0048] Also contemplated are malignancies located in the lung, colon, abdomen, bone, breast, digestive system, liver, pancreas, peritoneum, endocrine glands (adrenal, parathyroid, hypophysis, testicles, ovaries, thymus, thyroid), eye, head and neck, nervous system (central and peripheral), lymphatic system, pelvis, skin, soft tissue, spleen, thorax and, more particularly, childhood acute lymphoblastic leukemia, acute lymphoblastic leukemia, acute lymphocytic leukemia, acute myeloid leukemia, adrenocortical carcinoma, adult (primary) hepatocellular cancer, adult (primary) liver cancer, adult acute lymphocytic leukemia, adult acute myeloid leukemia, adult Hodgkin's disease, adult Hodgkin's lymphoma, adult lymphocytic leukemia, adult non-Hodgkin's lymphoma, adult primary liver cancer, adult soft tissue sarcoma, AIDS-related lymphoma, AIDS- related malignant tumors, anal cancer, astrocytoma, cancer of the biliary tract, cancer of the bladder, bone cancer, brain stem glioma, brain tumors, breast cancer, cancer of the renal pelvis and ureter, primary central nervous system lymphoma, central nervous system lymphoma, cerebellar astrocytoma, brain astrocytoma, cancer of the cervix, childhood (primary) hepatocellular cancer, childhood (primary) liver cancer, childhood acute lymphoblastic leukemia, childhood acute myeloid leukemia, childhood brain stem glioma, childhood cerebellar astrocytoma, childhood brain astrocytoma, childhood extracranial germ cell tumors, childhood Hodgkin's disease, childhood Hodgkin's lymphoma, childhood visual pathway and hypothalamic glioma, childhood lymphoblastic leukemia, childhood medulloblastoma, childhood non-Hodgkin's lymphoma, childhood supratentorial primitive neuroectodermal and pineal tumors, childhood primary liver cancer, childhood rhabdomyosarcoma, childhood soft tissue sarcoma, childhood visual pathway and hypothalamic glioma, chronic lymphocytic leukemia, chronic myeloid leukemia, cancer of the colon, cutaneous T-cell lymphoma, endocrine pancreatic islet cells carcinoma, endometrial cancer, ependymoma, epithelial cancer, Ewing's sarcoma and related tumors, cancer of the exocrine pancreas, extracranial germ cell tumor, extragonadal germ cell tumor, extrahepatic biliary tract cancer, cancer of the eye, breast cancer in women, Gaucher's disease, cancer of the gallbladder, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal tumors, germ cell tumors, gestational trophoblastic tumor, head and neck cancer, hepatocellular cancer, Hodgkin's disease, Hodgkin's lymphoma, hypergammaglobulinemia, hypopharyngeal cancer, intestinal cancers, intraocular melanoma, islet cell carcinoma, islet cell pancreatic cancer, Kaposi's sarcoma, cancer of kidney, cancer of the larynx, cancer of the lip and mouth, cancer of the liver, cancer of the lung, lymphoproliferative disorders, macroglobulinemia, breast cancer in men, malignant mesothelioma, malignant thymoma, medulloblastoma, melanoma, mesothelioma, occult primary metastatic squamous neck cancer, primary metastatic squamous neck cancer, metastatic squamous neck cancer, multiple myeloma, multiple myeloma / plasmatic cell neoplasia, myelodysplastic syndrome, myelogenous leukemia, myeloid leukemia, myeloproliferative disorders, paranasal sinus and nasal cavity cancer, nasopharyngeal cancer, neuroblastoma, nonHodgkin's lymphoma during pregnancy, non-melanoma skin cancer, non-small cell lung cancer, metastatic squamous neck cancer with occult primary, buccopharyngeal cancer, malignant fibrous histiocytoma, malignant fibrous osteosarcoma / histiocytoma of the bone, epithelial ovarian cancer, ovarian germ cell tumor, ovarian low malignant potential tumor, pancreatic cancer, paraproteinemias, purpura, parathyroid cancer, cancer of the penis, phaeochromocytoma, hypophysis tumor, neoplasia of plasmatic cells / multiple myeloma, primary central nervous system lymphoma, primary liver cancer, prostate cancer, rectal cancer, renal cell cancer, cancer of the renal pelvis and ureter, retinoblastoma, rhabdomyosarcoma, cancer of the salivary glands, sarcoidosis, sarcomas, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous neck cancer, stomach cancer, pineal and supratentorial primitive neuroectodermal tumors, T-cell lymphoma, testicular cancer, thymoma, thyroid cancer, transitional cell cancer of the renal pelvis and ureter, transitional renal pelvis and ureter cancer, trophoblastic tumors, cell cancer of the renal pelvis and ureter, cancer of the urethra, cancer of the uterus, uterine sarcoma, vaginal cancer, optic pathway and hypothalamic glioma, cancer of the vulva, Waldenstrom's macroglobulinemia, Wilms' tumor and any other hyperproliferative disease, as well as neoplasia, located in the system of a previously mentioned organ. A “chemotherapy agent,” “chemotherapeutic,” “anti-cancer agent,” or the like, refer to molecules that are recognized to aid in the treatment of a cancer. Contemplated examples include the following molecules or derivatives such as abemaciclib, abiraterone acetate, methotrexate, paclitaxel, adriamycin, acalabrutinib, brentuximab vedotin, ado-trastuzumab emtansine, aflibercept, afatinib, netupitant, palonosetron, imiquimod, aldesleukin, alectinib, alemtuzumab, pemetrexed disodium, copanlisib, melphalan, brigatinib, chlorambucil, amifostine, aminolevulinic acid, anastrozole, apalutamide, aprepitant, pamidronate disodium, exemestane, nelarabine, arsenic trioxide, ofatumumab, atezolizumab, bevacizumab, avelumab, axicabtagene ciloleucel, axitinib, azacitidine, carmustine, belinostat, bendamustine, inotuzumab ozogamicin, bevacizumab, bexarotene, bicalutamide, bleomycin, blinatumomab, bortezomib, bosutinib, brentuximab vedotin, brigatinib, busulfan, irinotecan, capecitabine, fluorouracil, carboplatin, carfilzomib, ceritinib, daunorubicin, cetuximab, cisplatin, cladribine, cyclophosphamide, clofarabine, cobimetinib, cabozantinib-S-malate, dactinomycin, crizotinib, ifosfamide, ramucirumab, cytarabine, dabrafenib, dacarbazine, decitabine, daratumumab, dasatinib, defibrotide, degarelix, denileukin diftitox, denosumab, dexamethasone, dexrazoxane, dinutuximab, docetaxel, doxorubicin, durvalumab, rasburicase, epirubicin, elotuzumab, oxaliplatin, eltrombopag olamine, enasidenib, enzalutamide, eribulin, vismodegib, erlotinib, etoposide, everolimus, raloxifene, toremifene, panobinostat, fulvestrant, letrozole, filgrastim, fludarabine, flutamide, pralatrexate, obinutuzumab, gefitinib, gemcitabine, gemtuzumab ozogamicin, glucarpidase, goserelin, propranolol, trastuzumab, topotecan, palbociclib, ibritumomab tiuxetan, ibrutinib, ponatinib, idarubicin, idelalisib, imatinib, talimogene laherparepvec, ipilimumab, romidepsin, ixabepilone, ixazomib, ruxolitinib, cabazitaxel, palifermin, pembrolizumab, ribociclib, tisagenlecleucel, lanreotide, lapatinib, olaratumab, lenalidomide, lenvatinib, leucovorin, leuprolide, lomustine, trifluridine, olaparib, vincristine, procarbazine, mechlorethamine, megestrol, trametinib, temozolomide, methylnaltrexone bromide, midostaurin, mitomycin C, mitoxantrone, plerixafor, vinorelbine, necitumumab, neratinib, sorafenib, nilutamide, nilotinib, niraparib, nivolumab, tamoxifen, romiplostim, sonidegib, omacetaxine, pegaspargase, ondansetron, osimertinib, panitumumab, pazopanib, interferon alfa-2b, pertuzumab, pomalidomide, mercaptopurine, regorafenib, rituximab, rolapitant, rucaparib, siltuximab, sunitinib, thioguanine, temsirolimus, thalidomide, thiotepa, trabectedin, valrubicin, vandetanib, vinblastine, vemurafenib, vorinostat, zoledronic acid, or combinations thereof such as cyclophosphamide, methotrexate, 5 -fluorouracil (CMF); doxorubicin, cyclophosphamide (AC); mustine, vincristine, procarbazine, prednisolone (MOPP); adriamycin, bleomycin, vinblastine, dacarbazine (ABVD); cyclophosphamide, doxorubicin, vincristine, prednisolone (CHOP); bleomycin, etoposide, cisplatin (BEP); epirubicin, cisplatin, 5- fluorouracil (ECF); epirubicin, cisplatin, capecitabine (ECX); methotrexate, vincristine, doxorubicin, cisplatin (MV AC). In certain embodiments, the chemotherapy agent is an anti-PD-1, anti-PD-Ll anti-CTLA4 antibody or combinations thereof, such as an anti-CTLA4 (e.g., ipilimumab, tremelimumab) and anti-PDl (e.g., nivolumab, pembrolizumab, cemiplimab) and anti-PD-Ll (e.g., atezolizumab, avelumab, durvalumab).

[0049] As used herein, the term “derivative” refers to a structurally similar compound that retains sufficient functional attributes of the identified analogue. The derivative may be structurally similar because it is lacking one or more atoms, substituted, a salt, in different hydration / oxidation states, or because one or more atoms within the molecule are switched, such as, but not limited to, replacing a oxygen atom with a sulfur atom, replacing an amino group with a hydroxyl group, replacing a nitrogen with a protonated carbon (CH) in an aromatic ring, replacing a bridging amino group (-NH-) with an oxy group (-O-), or vice versa. The derivative may be a prodrug. Derivatives may be prepared by any variety of synthetic methods or appropriate adaptations presented in synthetic or organic chemistry textbooks, such as those provide in March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Wiley, 6th Edition (2007) Michael B. Smith or Domino Reactions in Organic Synthesis, Wiley (2006) Lutz F. Tietze hereby incorporated by reference.

[0050] The term "prodrug" refers to an agent that is converted into a biologically active form in vivo. Prodrugs are often useful because, in some situations, they may be easier to administer than the parent compound. They may, for instance, be bioavailable by oral administration whereas the parent compound is not. The prodrug may also have improved solubility in pharmaceutical compositions over the parent drug. A prodrug may be converted into the parent drug by various mechanisms, including enzymatic processes and metabolic hydrolysis. Typical prodrugs are pharmaceutically acceptable esters. Prodrugs include compounds wherein a hydroxy, amino or mercapto group is bonded to any group that, when the prodrug of the active compound is administered to a subject, cleaves to form a free hydroxy, free amino or free mercapto group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate, and benzoate derivatives of an alcohol or acetamide, formamide and benzamide derivatives of an amine functional group in the active compound and the like.

[0051] The term "substituted" refers to a molecule wherein at least one hydrogen atom is replaced with a substituent. When substituted, one or more of the groups are "substituents." The molecule may be multiply substituted. In the case of an oxo substituent ("=O"), two hydrogen atoms are replaced. Example substituents within this context may include halogen, hydroxy, alkyl, alkoxy, nitro, cyano, oxo, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, -NRaRb, -NRaC(=O)Rb, -NRaC(=O)NRaNRb, -NRaC(=O)ORb, - NRaSCERb, -C(=O)Ra, -C(=O)ORa, -C(=O)NRaRb, -OC(=O)NRaRb, -ORa, -SRa, -SORa, - S(=O)2Ra, -OS(=O)2 I, and -S(=O)2ORa. Ra and Rb in this context may be the same or different and independently hydrogen, halogen hydroxyl, alkyl, alkoxy, alkyl, amino, alkylamino, dialkylamino, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl.

[0052] Methods of Use

[0053] This disclosure relates to methods of treating cancer comprising administering an effective amount of an aminoadipate-semialdehyde synthase inhibitor to a subject in need thereof. In certain embodiments, the aminoadipate-semialdehyde synthase inhibitor is N-benzyl-N,N- dimethylhexadecan-l-aminium chloride (cetalkonium chloride) or salts thereof. In certain embodiments, the subject is a human patient.

[0054] In certain embodiments, methods disclosed herein further comprise administering an additional anti-cancer agent in combination with the aminoadipate-semialdehyde synthase inhibitor such as N-benzyl-N,N-dimethylhexadecan-l-aminium chloride (cetalkonium chloride) salts thereof. In certain embodiments, the aminoadipate-semialdehyde synthase inhibitor is a N- benzyl-N,N-dimethylhexadecan-l-aminium salt, prodrug, metabolite, or derivative thereof.

[0055] In certain embodiments, the aminoadipate-semialdehyde synthase inhibitor is an antibody, small molecule compound, peptide, or siRNA.

[0056] In certain embodiments, the cancer is selected from a solid cancer, advanced cancer, metastatic cancer, and / or hematological cancer. In certain embodiments, the solid cancer has metastasized or is a locally advanced cancer. In certain embodiments, the solid cancer is selected from liver cancer, bladder cancer, lung cancer, ovarian cancer, breast cancer, colon cancer, rectal cancer, endometrial cancer, pancreatic cancer, kidney cancer, prostate cancer, thyroid cancer, or brain cancer. In certain embodiments, hematological cancer is leukemia, lymphoma, or multiple myeloma.

[0057] In certain embodiments, methods further comprise administering an additional anti-cancer agent.

[0058] In certain embodiments, this disclosure relates to methods of treating cancer comprising administering an effective amount of an aminoadipate-semialdehyde synthase inhibitor to a subject in need thereof. In certain embodiments, this disclosure relates to methods of detecting amounts of aminoadipate-semialdehyde synthase in a sample to determine or monitor whether the subject is sensitive or resistant to an aminoadipate-semialdehyde synthase inhibitor or combination of chemotherapy agents comprising the same.

[0059] In certain embodiments, this disclosure relates to methods of detecting amounts of aminoadipate-semialdehyde synthase in a sample to determine whether the subject is sensitive or resistant or is likely to develop resistance to a chemotherapy agent or combination of chemotherapy agents comprising the same. In certain embodiments, the sample is derived from tumor cells or cell-free DNA contained in a blood sample of a subject diagnosed with cancer.

[0060] The precise therapeutically effective amount of an aminoadipate-semialdehyde synthase inhibitor disclosed herein will depend on a number of factors. These variables determine what dose of compound needs to be administered in a sufficient percentage and for a sufficient amount of time to have the desired effect on the condition being treated (e g., cancer). The amount of compound administered will also depend on factors related to patients and disease including, but not limited to, the following: the age, weight, concomitant medications, and medical condition of the subject being treated, the precise condition requiring treatment and its severity, the nature of the formulation, and the route of administration. Ultimately, the dose will be at the discretion of the attendant physician or veterinarian. Typically, the compound disclosed herein will be given for treatment in the range of 0.01 to 30 mg / kg body weight of recipient (mammal) per day or per dose or per cycle of treatment and more usually in the range of 0.1 to 10 mg / kg body weight per day or per dose or per cycle of treatment. Thus, for an adult human being treated for a condition, the actual amount per day or per dose or per cycle of treatment would usually be from 1 to 2000 mg and this amount may be given in a single or multiple doses per day or per dose or per cycle of treatment. The full spectrum of dosing regimens may be employed ranging from continuous dosing (with daily doses) to intermittent dosing.

[0061] In the above-described methods of treatment and uses, aminoadipate-semialdehyde synthase inhibitor herein may be employed alone, in combination with one or more other compounds disclosed herein or in combination with other therapeutic methods or agents. In certain embodiments, methods of treating are done in combination with administering other chemotherapeutic, biologic, hormonal, antibody, and supportive care agents. Such methods may also be performed in combination with surgical therapy, radiation, and imaging methods. Supportive care agents include analgesics, anti-emetics and agents used to treat hematologic side effects such as neutropenia. Analgesics are well known in the art. Anti-emetics include but are not limited to 5HT3 antagonists such as ondansetron, granisetron, dolasetron, palonosetron and the like; prochlorperazine; metoclopramide; diphenhydramine; promethazine; dexamethasone; lorazepam; haloperidol; dronabinol; olanzapine; and neurokinin- 1 antagonists such as aprepitant, fosaprepitant and casopitant administered alone or in various combinations.

[0062] Combination therapies according to the disclosure thus comprise the administration of aminoadipate-semialdehyde synthase inhibitors disclosed herein and the use of at least one other treatment method. In certain embodiments, combination therapies according to the disclosure comprise the administration of aminoadipate-semialdehyde synthase inhibitors disclosed herein and surgical therapy. In certain embodiments, combination therapies according to the disclosure comprise the administration of aminoadipate-semialdehyde synthase inhibitors disclosed herein and radiotherapy. In certain embodiments, combination therapies according to the disclosure comprise the administration of aminoadipate-semialdehyde synthase inhibitors disclosed herein and at least one supportive care agent (e.g., at least one anti-emetic agent). In certain embodiments, combination therapies according to the present disclosure comprise the administration of aminoadipate-semialdehyde synthase inhibitors disclosed herein and at least one other chemotherapeutic agent.

[0063] In certain embodiments, the present disclosure provides the methods of treatment and uses as described above, which comprise administering an aminoadipate-semialdehyde synthase inhibitor disclosed herein together with at least one other chemotherapeutic agent. In certain embodiments, the disclosure also provides methods of treatment and uses as described above, which comprise administering an aminoadipate-semialdehyde synthase inhibitor disclosed herein together with at least one supportive care agent (e.g., anti-emetic agent).

[0064] In certain embodiments, the aminoadipate-semialdehyde synthase inhibitors disclosed herein and at least one additional anti-cancer or supportive care therapy may be employed in combination concomitantly or sequentially in any therapeutically appropriate combination. The administration of aminoadipate-semialdehyde synthase inhibitors disclosed herein with one or more other anti-cancer agents may be in combination in accordance with the disclosure by administration concomitantly in one unitary pharmaceutical composition including both or all compounds or two or more separate pharmaceutical compositions each including one or more of the compounds. The components of the combination may be administered separately in a sequential manner wherein one active ingredient is administered first and the other(s) second or vice versa. Such sequential administration may be close in time or remote in time.

[0065] Typically, any chemotherapeutic agent that has activity against a susceptible cancer being treated may be utilized in combination with aminoadipate-semialdehyde synthase inhibitors herein, provided that the agent is clinically compatible with a therapy employing a compound disclosed herein. Typical anti-cancer agents useful in the present disclosure include, but are not limited to: alkylating agents, anti-metabolites, antitumor antibiotics, antimitotic agents, topoisomerase I and II inhibitors, hormones and hormonal analogues; retinoids, signal transduction pathway inhibitors including inhibitors of cell growth or growth factor function, angiogenesis inhibitors, and serine / threonine or other kinase inhibitors; cyclin dependent kinase inhibitors; antisense therapies and immunotherapeutic agents, including monoclonals, vaccines or other biological agents.

[0066] Alkylating agents are non-phase specific anti-cancer agents and strong electrophiles. Typically, alkylating agents form covalent linkages, by alkylation, to DNA through nucleophilic moieties of the DNA molecule such as phosphate, amino, and hydroxyl groups. Such alkylation disrupts nucleic acid function leading to cell death. Alkylating agents may be employed in combination with the aminoadipate-semialdehyde synthase inhibitors disclosed herein in the compositions and methods described above. Examples of alkylating agents include but are not limited to nitrogen mustards such as cyclophosphamides, temozolomide, melphalan, and chlorambucil; oxazaphosphorines; alkyl sulfonates such as busulfan; nitrosoureas such as carmustine; triazenes such as dacarbazine; and platinum coordination complexes such as cisplatin, oxaliplatin and carboplatin.

[0067] Antimetabolite neoplastic agents may be employed in combination with the aminoadipatesemialdehyde synthase inhibitors disclosed herein in the compositions and methods described above. Examples of antimetabolite anti-cancer agents include but are not limited to purine and pyrimidine analogues and anti-folate compounds, and more specifically, hydroxyurea, cytosine, arabinoside, raltitrexed, tegafur, fluorouracil (e.g., 5FU), methotrexate, cytarabine, mercaptopurine and thioguanine.

[0068] Antitumor antibiotic agents are non-phase specific agents, which bind to or intercalate with DNA. Typically, such action disrupts ordinary function of the nucleic acids, leading to cell death. Antitumor antibiotics may be employed in combination with the aminoadipate-semialdehyde synthase inhibitors disclosed herein in the compositions and methods described above. Examples of antitumor antibiotic agents include, but are not limited to, actinomycins such as dactinomycin; anthracyclines such as daunorubicin, doxorubicin, idarubicin, epirubicin and mitoxantrone; mitomycin C and bleomycins.

[0069] Antimicrotubule or antimitotic agents are phase specific agents active against the microtubules of tumor cells during M or the mitosis phase of the cell cycle. Antimitotic agents may be employed in combination with the aminoadipate-semialdehyde synthase inhibitors disclosed herein in the compositions and methods described above. Examples of antimitotic agents include, but are not limited to, diterpenoids, vinca alkaloids, and polo-like kinase (Plk) inhibitors. Examples of diterpenoids include, but are not limited to, paclitaxel and its analog docetaxel. Examples of vinca alkaloids include, but are not limited to, vinblastine, vincristine, vindesine and vinorelbine.

[0070] Topoisomerase inhibitors include inhibitors of Topoisomerase II and inhibitors of Topoisomerase I. Topoisomerase II inhibitors, such as epipodophyllotoxins, are anti-cancer agents that typically affect cells in the S and G2 phases of the cell cycle by forming a ternary complex with topoisomerase II and DNA, causing DNA strand breaks. The strand breaks accumulate, and cell death follows. Examples of epipodophyllotoxins include, but are not limited to, etoposide and teniposide. Camptothecins, including camptothecin and camptothecin derivatives, are available or under development as Topoisomerase I inhibitors. Examples of camptothecins include, but are not limited to amsacrine, irinotecan, topotecan, and the various optical forms of 7-(4- methylpiperazino-methylene)-10,l 1 -ethyl enedioxy-20-camptothecin. Topoisomerase inhibitors may be employed in combination with the aminoadipate-semialdehyde synthase inhibitors disclosed herein in the compositions and methods described above.

[0071] Hormones and hormonal analogues are useful compounds for treating cancers in which there is a relationship between the hormone(s) and growth and / or lack of growth of the cancer. Antitumor hormones and hormonal analogues may be employed in combination with the aminoadipate-semialdehyde synthase inhibitors disclosed herein in the compositions and methods described above. Examples of hormones and hormonal analogues believed to be useful in the treatment of cancers include, but are not limited to antiestrogens, such as tamoxifen, toremifene, raloxifene, fulvestrant, idoxifene and droloxifene; anti-androgens; such as flutamide, nilutamide, bicalutamide and cyproterone acetate; adrenocorticosteroids such as prednisone and prednisolone; aminoglutethimide and other aromatase inhibitors such as anastrozole, letrozole, vorozole, and exemestane; progestins such as megestrol acetate; 5 alpha-reductase inhibitors such as finasteride and dutasteride; and gonadotropin-releasing hormones (GnRH) and analogues thereof, such as Luteinizing Horm one-releasing Hormone (LHRH) agonists and antagonists such as goserelin leuprolide, leuprorelin and buserelin.

[0072] Examples of specific retinoids that may be used in combination with the aminoadipate- semialdehyde synthase inhibitors disclosed herein include: retinoic acid; all-trans-retinoic acid ("ATRA" also known as "tretinoin"); tamibarotene ("Am80"); 9-cis-retinoic acid ((2E,4E,6Z,8E)- 3,7-Dimethyl-9-(2,6,6-trimethylcyclohex-l-enyl)nona-2,4,6,- 8-tetraenoic Acid) (also known as "9-cis-Tretinoin") (available from Sigma); Isotretinoin ((2Z,4E,6E,8E)-3,7-dimethyl-9-(2,6,6- trimethyl-l-cyclohexenyl)nona-2,4,6,- 8-tetraenoic acid) (also known as " 13-cis-retinoic acid") (ACCUTANE1M); Am580 (4-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphtamido) benzoic acid), See, M. Gianni, Blood 1996 87(4): 1520-1531; TTNPB (4-[E-2-(5, 6,7, 8-Tetrahydro-5, 5,8,8- tetramethyl-2-naphthalenyl)-l-propeny- l]benzoic acid) (also known as "Ro 13-7410") See, M. F. Boehm et al. J. Med. Chem. 1994 37:2930 and R. P. Bissonnette et al., Mol. Cell. Biol. 1995 15:5576; and BMS753 (4-[[(2,3-dihydro-l,l,3,3-tetramethyl-2-oxo-lH-inden-5- yl)carbonyl]amino]- benzoic acid) See, U.S. Pat. No. 6,184,256.

[0073] Several inhibitors of growth factor receptors are contemplated including ligand antagonists, antibodies, tyrosine kinase inhibitors, anti-sense oligonucleotides and aptamers. Any of these growth factor receptor inhibitors may be employed in combination with aminoadipate- semialdehyde synthase inhibitors disclosed herein in any of the compositions and methods / uses described herein. Trastuzumab is an example of an anti-erbB2 antibody inhibitor of growth factor function. One example of an anti-erbBl antibody inhibitor of growth factor function is cetuximab. Bevacizumab is an example of a monoclonal antibody directed against VEGFR. Examples of small molecule inhibitors of epidermal growth factor receptors include but are not limited to lapatinib and erlotinib. Imatinib is one example of a PDGFR inhibitor. Examples of VEGFR inhibitors include pazopanib, sunitinib and sorafenib.

[0074] In certain embodiments, aminoadipate-semialdehyde synthase inhibitors disclosed herein can be used and formulated in combination with the anti-cancer agents that are PD-1 antibodies such as nivolumab, pembrolizumab, pidilizumab, atezolizumab or CTLA-4 antibodies such as ipilimumab and tremelimumab.

[0075] The pharmaceutical compositions comprising aminoadipate-semialdehyde synthase inhibitors can be administered to subjects either orally, rectally, parenterally (intravenously, intramuscularly, or subcutaneously), intracistemally, intravaginally, intraperitoneally, intravesically, locally (powders, ointments, or drops), or as a buccal or nasal spray. Pharmaceutically acceptable salts, solvates and hydrates of the compounds listed are also useful in the method of the disclosure and in pharmaceutical compositions of the disclosure.

[0076] Pharmaceutical compositions and kits

[0077] In certain embodiments, this disclosure relates to pharmaceutical composition comprising an aminoadipate-semialdehyde synthase inhibitor and a pharmaceutically acceptable excipient for uses disclosed herein. In certain embodiments, the aminoadipate-semialdehyde synthase inhibitor is a N-benzyl-N,N-dimethylhexadecan-l-aminium salt, prodrug, metabolite, or derivative thereof. In certain embodiments, the aminoadipate-semialdehyde synthase inhibitor is N-benzyl-N,N- dimethylhexadecan-l-aminium chloride (cetalkonium chloride).

[0078] In certain embodiments, the pharmaceutical composition is in the form of a lotion, gel, cream, pill, tablet, capsule, microparticle, or gel capsule. In certain embodiments, the pharmaceutical composition is in the form of a sterilized pH buffered aqueous salt solution or a saline phosphate buffer between a pH of 6 to 8, optionally comprising a saccharide or polysaccharide. In certain embodiments, this disclosure relates to medicaments comprising an aminoadipate-semialdehyde synthase inhibitor for use in treating cancer as provided herein. In certain embodiments, this disclosure relates to medicaments comprising an aminoadipate- semialdehyde synthase inhibitor for use in treating cancer as provided herein.

[0079] In certain embodiments, this disclosure relates to pharmaceutical compositions and kits comprising aminoadipate-semialdehyde synthase inhibitors disclosed herein. In certain embodiments, this disclosure relates to the production of a medicament comprising aminoadipate- semialdehyde synthase inhibitors disclosed herein for therapeutic uses reported herein.

[0080] In certain embodiments, this disclosure relates to pharmaceutical compositions comprising aminoadipate-semialdehyde synthase inhibitors disclosed herein. In certain embodiments, the pharmaceutical composition optionally comprises a pharmaceutical carrier, and the pharmaceutical composition optionally comprises further therapeutic agents, anticancer agents, anti-inflammatory agents, etc. In certain embodiments, a pharmaceutical composition is in the form of a liquid comprising pH buffering agents and optionally salts and / or saccharide or polysaccharide.

[0081] In certain embodiments, this disclosure contemplates an intravenous formulation with pH buffering agents and tonicity in a range representing physiological values (neutral of about pH 7 to 8) or for bolus administration, e.g., containing normal saline or dextrose optionally containing pH buffering agents. In certain embodiments, the pharmaceutical composition is in the form of a sterilized pH buffered aqueous salt solution or a saline phosphate buffer between a of about pH of 6 to 8, optionally comprising a saccharide or polysaccharide.

[0082] In certain embodiments, this disclosure relates to pharmaceutical compositions comprising aminoadipate-semialdehyde synthase inhibitors disclosed herein and a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutically acceptable excipient is selected from lactose, sucrose, mannitol, triethyl citrate, dextrose, cellulose, methyl cellulose, ethyl cellulose, hydroxyl propyl cellulose, hydroxypropyl methylcellulose, carboxymethylcellulose, croscarmellose sodium, polyvinyl N-pyrrolidone, crospovidone, ethyl cellulose, povidone, methyl and ethyl acrylate copolymer, polyethylene glycol, fatty acid esters of sorbitol, lauryl sulfate, gelatin, glycerin, glyceryl monooleate, silicon dioxide, titanium dioxide, talc, corn starch, carnauba wax, stearic acid, sorbic acid, magnesium stearate, calcium stearate, castor oil, mineral oil, calcium phosphate, starch, carboxymethyl ether of starch, iron oxide, triacetin, acacia gum, esters, or salts thereof.

[0083] Compositions suitable for parenteral injection may comprise physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Examples of suitable aqueous and nonaqueous carriers, diluents solvents or vehicles include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, and the like), suitable mixtures thereof, vegetable (such as olive oil, sesame oil) and injectable organic esters such as ethyl oleate.

[0084] These compositions may also contain preserving, emulsifying, and dispensing agents. Prevention of the action of microorganisms may be controlled by addition of any of various antibacterial and antifungal agents, example, parabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agents, for example sugars, sodium chloride, and the like. Prolonged absorption of the injectable pharmaceutical form may be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0085] Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, capsules, gel capsules, and pills. In addition to the phosphate membrane nanodiscs conjugated to therapeutic agents as reported herein, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, in particular, cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan or mixtures of these substances, and the like.

[0086] In certain embodiments, the pharmaceutical compositions comprising aminoadipatesemialdehyde synthase inhibitors disclosed herein can be administered to subjects either orally, parenterally (intravenously, intramuscularly, or subcutaneously), intracisternally, intraperitoneally, intravesically, locally (powders, ointments, or drops), intravaginally, as a buccal or nasal spray, topically to the skin, or rectally.

[0087] In certain embodiments, the pharmaceutical compositions are in a form for inhalation. In certain embodiments, the pharmaceutical composition comprises aminoadipate-semialdehyde synthase inhibitors disclosed herein and a propellant. In certain embodiments, an aerosolizing propellant is compressed air, ethanol, nitrogen, carbon dioxide, nitrous oxide, hydrofluoroalkanes (HF As), or combinations thereof.

[0088] In certain embodiments, the disclosure contemplates a pressurized or unpressurized container comprising aminoadipate-semialdehyde synthase inhibitors disclosed herein. In certain embodiments, the container is a manual pump spray, inhaler, meter-dosed inhaler, dry powder inhaler, nebulizer, vibrating mesh nebulizer, jet nebulizer, or ultrasonic wave nebulizer.

[0089] In certain embodiments, this disclosure contemplates kits comprising pharmaceutical compositions comprising aminoadipate-semialdehyde synthase inhibitors disclosed herein and optionally another therapeutic agent / anti cancer agent in same or separate pharmaceutical composition or container. The kits may contain a transfer device such a needle, syringe, cannula, capillary tube, pipette, or pipette tip. In certain embodiments, the agents may be contained in a storage container, sealed, or unsealed, such a vial, bottle, ampule, blister pack, or box. In certain embodiments, the kit further comprises written instructions for using the agents for treating and / or preventing cancer or other disease or condition in a subject.

[0090] In certain embodiments, this disclosure relates to uses of aminoadipate-semialdehyde synthase inhibitors disclosed herein in the production of a medicament for treating diseases of conditions disclosed herein.

[0091] Dosing is dependent on severity and responsiveness of the disease state to be treated, and the course of treatment lasting from several days to several months, or until a cure is effected or a diminution of the disease state is achieved. Optimal dosing schedules can be calculated from measurements of drug accumulation in the body of the patient. Optimum dosages may vary depending on the relative potency of individual oligonucleotides. Generally, it can be estimated based on amounts found to be effective in in vitro and in vivo animal models. In general, dosage is from 0.01 pg to 1 g per kg of body weight, and may be given once or more daily, weekly, monthly, or yearly, or even once every 2 to 10 years or by continuous infusion for hours up to several months. The repetition rates for dosing can be estimated based on measured residence times and concentrations of the drug in bodily fluids or tissues. Following successful treatment, it may be desirable to have the patient undergo maintenance therapy to prevent the recurrence of the disease state. Aminoadipate-semialdehyde synthase inhibitors and methods of treating cancer

[0092] Metastases are the cause of a large majority of cancer deaths. Growing evidence suggests that metabolic difference exists between primary tumor and metastasized tumor. While primary cancer cells often prefer aerobic glycolysis, mitochondrial factors are reported to be upregulated in metastasized tumor cells. Understanding the essential metabolic pathway that promotes metastasis is vitally important for successful clinical outcome of cancer patients with metastatic disease.

[0093] To identify a mitochondrial factor critical for cancer metastasis, an unbiased RNAi-based screen was performed using a customized RNAi library targeting mitochondrial enzymes. Screen using various types of cancer cells (lung, head / neck, breast, ovarian cancers) revealed that aminoadipate-semialdehyde synthase (AASS) is a common factor for invasion. In vitro and in vivo studies using AASS variants demonstrated that the activity of AASS is required for cancer cell invasion and metastasis. Targeting enzymes in the saccharopine pathway revealed 2-aminoadipate (2-AAA), a metabolic intermediate, as an important factor for cell invasion. Experiments evaluating metabolic, genomic, and transcriptomics, indicate that AASS promotes cancer cell invasion and metastasis by promoting redox homeostasis and epithelial-mesenchymal transition (EMT). Furthermore, AASS upregulation positively correlates with metastatic progression in tumor tissues from cancer patients, suggesting AASS as a promising prognostic marker and a therapeutic target to halt metastasis.

[0094] Screening small molecule compounds, cetalkonium chloride (CKC) was identified as potent AASS inhibitor and an anti-metastasis agent. Treatment of CKC in various cancer cell lines (IC50 4-5.2 uM) and mice dramatically reduced invasive potential in vitro and metastasis in vivo. The effect of CKC on invasion inhibition is specifically through AASS inhibition since the drug was not effective when cells lack AASS. Furthermore, both chronic and acute injection of CKC did not significantly alter hematopoietic and histological properties in mice suggesting limited toxicity of CKC in vivo.

[0095] Inhibition of Aminoadipate-semialdehyde synthase (AASS) in the saccharopine pathway

[0096] In vitro and in vivo xenograft studies using AASS variants revealed that the activity of AASS is commonly important for cancer cell invasion and metastasis in various solid tumors including lung, head and neck, breast, and ovarian cancers. Target downregulation of each of the enzymes in the saccharopine pathway showed that a metabolic intermediate, 2-aminoadipate (2- AAA), is the factor that provides pro-invasive advantage to cancer cells. To glean mechanistic insight into how AASS promotes metastasis, metabolic, genomic, and transcriptomics approaches were performed. A series of metabolic assays demonstrated that AASS contributes to pro-invasive potential by managing redox balance in cells. Transcriptome sequencing and gene set enrichment analysis (GSEA) revealed that AASS loss results in decrease of peroxisomal genes and a redox regulating factor, thioredoxin domain-containing protein 5 (TXNDC5). Moreover, the spliced form of x-box binding protein 1 (sXBPl), an activated transcription factor of TXNDC5, was reduced whereas un-spliced XBP1 was elevated in cells lacking AASS, suggesting a possible connection among AASS, sXBPl, and TXNDC5. In addition, through kinase inhibitor profding and transcription factor activation profiling, hepatocyte growth factor (HGF) / c-MET was identified as an upstream kinase of AASS and SATB1 as a potential AASS transcription factor that may regulate AASS activation and expression during cancer cell invasion and metastasis. Experiments indicate that AASS expression positively correlates with metastatic progression in a panel of cancer cell lines including lung cancer and head / neck cancer, and a small cohort of patient tumor tissues. This suggests that AASS may be a promising therapeutic target to overcome metastasis.

[0097] Screening to identify clinically approved small molecule compounds

[0098] Cetalkonium chloride was identified as an AASS inhibitor. Cetalkonium chloride effectively inhibits AASS activity in cancer cells and attenuates cell invasion. AASS and its metabolic intermediate 2-AAA contribute to metastasis through potential effectors and regulators including XBP1, TXNDC5, c-MET and SATB1; AASS represents an attractive therapeutic target in metastatic cancer treatment.

Claims

CLAIMS1. A method of treating cancer comprising administering an effective amount of an aminoadipate-semialdehyde synthase inhibitor to a subject in need thereof.

2. The method of claim 1, wherein the aminoadipate-semialdehyde synthase inhibitor is a N- benzyl-N,N-dimethylhexadecan-l-aminium salt, prodrug, metabolite, or derivative thereof.

3. The method of claim 1, wherein the aminoadipate-semialdehyde synthase inhibitor is N- benzyl-N,N-dimethylhexadecan-l-aminium chloride (cetalkonium chloride).

4. The method of claim 1, wherein the aminoadipate-semialdehyde synthase inhibitor is an antibody, small molecule compound, peptide, or siRNA.

5. The method of claim 1, wherein the cancer is selected from a solid or hematological cancer.

6. The method of claim 5, wherein the hematological cancer is leukemia, lymphoma, or multiple myeloma.

7. The method of claim 5 wherein the solid cancer is selected from live cancer, bladder cancer, lung cancer, breast cancer, colon cancer, rectal cancer, endometrial cancer, pancreatic cancer, kidney cancer, prostate cancer, thyroid cancer, or brain cancer.

8. The method of claim 5 wherein the solid cancer has metastasized.

9. The method of claim 1, wherein the method further comprises administering an additional anti-cancer agent.

10. A pharmaceutical composition comprising aminoadipate-semialdehyde synthase inhibitor and a pharmaceutically acceptable excipient.

11. The pharmaceutical composition of claim 10 wherein the aminoadipate-semialdehyde synthase inhibitor is a N-benzyl-N,N-dimethylhexadecan-l-aminium salt, prodrug, metabolite, or derivative thereof12. The pharmaceutical composition of claim 10 wherein the aminoadipate-semialdehyde synthase inhibitor is N-benzyl-N,N-dimethylhexadecan-l-aminium chloride (cetalkonium chloride).

13. The pharmaceutical composition of claim 10 in the form of a lotion, gel, cream, pill, tablet, capsule, or gel capsule.

14. The pharmaceutical composition of claim 10 in the form of a sterilized pH buffered aqueous salt solution or a saline phosphate buffer between a pH of 6 to 8, optionally comprising a saccharide or polysaccharide.

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